I am planning to set up new experiments for a student´s lab course in our master´s degree course in biotechnology. We have an automated inverted fluorescence microscope, Olympus IX81, which nobody has been using in the last years and which I would like to integrate in the course.
As I have very little experience in this field, I was wondering which experiments would be easiest to start with?
We have a small cell culture lab - so we could grow e.g. adherent cells in culture flasks etc., but we don´t have a microtom for making tissue section etc.
Looking forward to any suggestions! Thanks!
If you can grow cells in culture, you probably don't need to do anything else at all. Just make sure you have an appropriate holder to mount the culture plate on the microscope stage, and use the transmitted light channel to image the cells if they don't have any fluorescent label. Then let the students watch the cells die due to being exposed to room temperature :) Or, if it has an incubator chamber, maintain the appropriate temperature and watch them live...
Hi Agnes,
As stated by Sebastian, you should use microscopy as an illustration of scientific questions. Microscopy isn't interesting by itself, it's a technique that allows to answer scientific questions.
You are talking to master's students, tell them science stories. Tell them about your science.
What are your research fields ? Howard microscopy coule give answers to your questions ?
If you just want an easy experiment to show students, you can by red onions or red cabbage, and show them the cell wall ans the stained vacuoles, then add sucrose or glycerol and see them plasmolyse in few seconds, them get turgescent again within minutes.
You can also see the stomata movements on lettuce leaves.
For fluorescence observations plant tissues are fabulous. You can see the blue fluorescence of xylem under UV, the flavonoids and chlorophyls under blue lights (with a long pass filter). Then add fluorescein di-acetate and show the cytosol stain, while raw fluorescein doesn't penetrate cells. You can also use dihydrorhodamine123 that stains in green mitochondria, and watch them move around the cell.
You can add DAPI and observe that the cells aren't stained, then add a depolarizing drug and see the mitochondria ans nuclei being stained.
All these are easy and funny, but I'm not sur they are enough to amaze master's students.
Cheers,
Oliv
Thanks for your answers!
The main focus on the lab course is to give the students a small insight into the matter of fluorescence microscopy. We have probably 4-6- hours max for that, as there are other topics in this lab course as well.Therefore we won´t have time to do some GFP-fusion constructs and I think I would also have a hard time agrueing that we will need to money for such a construct just for a demo ;-)
Fluorescence microscopy per se doesn´t fit directly into any our research topics, but we do have a cell culture lab, so we could cultivate some endothelial cell and stain them with DAPI and for actin etc. But I am not sure how difficult it is to fix the cells and do the staining ...
As for the plant tissue: do you need a microtom to make a section thin enough for microscopy? If not, that might be an intersting topic!
Thanks,
Agnes
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